title: Identification of a second cytotoxic protein produced by Bacillus thuringiensis A1470.
abstract: Bacillus thuringiensis A1470 produces multiple proteins with similar molecular masses (~30 kDa) with cytotoxicity against human cell lines. One that was previously identified, parasporin-4, is a β-pore-forming toxin. The N-terminal sequence of a second cytotoxic protein was identical to a partial sequence of parasporin-2 produced by B. thuringiensis A1547. PCR was performed on total plasmid DNA from A1470 by using primers for parasporin-2 to amplify a gene which was then cloned. The cloned gene differed from A1547 parasporin-2 by 8 bp and the predicted protein differed by four amino acids. The gene was expressed in Escherichia coli, and the cytotoxic activities of the recombinant protein against four human cell lines (MOLT-4, Jurkat, HeLa, and HepG2) were similar to those of A1547 parasporin-2. We then confirmed that the A1470 strain simultaneously produces parasporin-2 and parasporin-4.
authors.affiliation: Biotechnology and Food Research Institute, Fukuoka Industrial Technology Center, 1465-5 Aikawa-machi, Kurume, Fukuoka, 839-0861, Japan, sokumura@fitc.pref.fukuoka.jp.
authors.forename: Shiro
authors.initials: S
authors.lastname: Okumura
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Ishikawa
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Saitoh
authors.forename: Tetsuyuki
authors.initials: T
authors.lastname: Akao
authors.forename: Eiichi
authors.initials: E
authors.lastname: Mizuki
mesh_terms: D001413:Bacillus thuringiensis / Q000235:genetics / Q000378:metabolism*; D001427:Bacterial Toxins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D003001:Cloning, Molecular; D003603:Cytotoxins / Q000235:genetics / Q000378:metabolism*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000633:toxicity; D017422:Sequence Analysis, DNA
chemical_list: D001427:Bacterial Toxins; D003603:Cytotoxins; D004269:DNA, Bacterial; D011994:Recombinant Proteins